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[Ectothiorhodosinus mongolicum gen. nov., sp. nov.,--a new purple sulfur bacterium from soda lake in Mongolia].

A new purple sulfur bacterium (strain M9) was isolated from the steppe soda Lake Dzun Uldziin Nur (pH 9.4; mineralization, 3.3%) situated in southeastern Mongolia. Individual cells appear as vibrios 0.3-0.5 x 0.7-1 micron in size. The dividing cells often do not separate from each other, forming an almost closed ring. The internal photosynthetic membranes are represented by concentric lamellae lining the cell wall. Photosynthetic pigments are bacteriochlorophyll a and carotenoids of the spirilloxanthin series. The main carotenoid (> 96%) is spirilloxanthin. Two typical light-harvesting complexes (LH1 and LH2) are present in the membranes in a 1:1 ratio. The bacterium is an anaerobe and facultative photoorganoheterotroph. Photolithoautotrophic growth on sulfide is scarce. Thiosulfate is utilized as an electron donor only in the presence of organic matter. Globules of elemental sulfur are formed as an intermediary product of sulfide and thiosulfate oxidation and are deposited outside the cells. The end product of oxidation is sulfate. In the presence of sulfide and carbonates, acetate, lactate, malate, pyruvate, propionate, succinate, and fumarate are used as the additional sources of carbon in anoxygenic photosynthesis. Vitamin are not required. The bacterium is an alkaliphile the pH optimum is at 8.3-9.1, the pH range is 7.6-10.1. The optimum NaCl concentration in the medium is 1 to 7%; the range is 0.5 to 0.9%. The optimum carbonate content in the medium is 2%; the range is 1 to 10%. The best growth occurs at 30-35 degrees C. The DNA G + C content is 57.5 mol%. According to the results of analysis of the 16S rRNA gene sequences, the new isolate M9 belongs to the phylogenetic cluster containing representatives of the family Ectothiorhodospiraceae within the class "Gammaproteobacteria." In this class, the new isolate forms a new branch, which occupies an intermediate position between the representatives of the genera Ectothiorhodospira and Thiorhodospira. Based on the phenotypic and genetic characteristics, the new purple sulfurbacterium was assigned to a new species of a new genus of the family Ectothiorhodospiraceae, Ectothiorhodosinus mongolicum gen. nov., sp. nov.

Anaerobiosis↗

[Effect of cultivation conditions on the growth and activities of sulfur metabolism enzymes and carboxylases of Sulfobacillus thermosulfidooxidans subsp. asporogenes strain 41].

The moderately thermophilic acidophilic bacterium Sulfobacillus thermosulfidooxidans subsp. asporogenes strain 41 is capable of utilizing sulfides of gold-arsenic concentrate and elemental sulfur as a source of energy. The growth in the presence of S0 under auto- or mixotrophic conditions was less stable compared with the media containing iron monoxide. The enzymes involved in oxidation of sulfur inorganic compounds--thiosulfate-oxidizing enzyme, tetrathionate hydrolase, rhodonase, adenylyl sulfate reductase, sulfite oxidase, and sulfur oxygenase--were discovered in the cells of Sulfobacillus grown in the mineral medium containing 0.02% yeast extract and either sulfur or iron monoxide and thiosulfate. Cell-free extracts of the cultures grown in the medium with sulfur under auto- or mixotrophic conditions displayed activity of the key enzyme of the Calvin cycle--ribulose bisphosphate carboxylase--and several other enzymes involved in heterotrophic fixation of carbonic acid. Activities of carboxylases depended on the composition of cultivation media.

Culture Media↗

New crystalline derivatives of bovine liver rhodanese.

Mitochondrial bovine liver rhodanese (thiosulfate:cyanide sulfurtransferase) has been crystallized in the form deprived of the transferable sulfur. The essential condition for crystallization was the removal of oxygen. Crystals of the sulfur-free enzyme are isomorphous with the previously characterized crystals of the sulfur-substituted enzyme. The new crystal species can react with either thiosulfate or selenosulfate to form the catalytic intermediate and, subsequently, with cyanide to form the corresponding product. Furthermore, the enzyme active site can be alkylated by iodoacetic acid.

Animals↗

Properties of an Escherichia coli rhodanese.

A rhodanese enzyme of less than 20,000 molecular weight has been purified from Escherichia coli. The enzyme is accessible to substrates upon addition of whole cells to standard assay mixtures. This rhodanese has a Stokes radius of 17 A which for a globular protein corresponds to a molecular weight close to 14,000. It undergoes autoxidation to a polymeric form which is probably an inert dimer. Enzyme inactivated by oxidation can be reactivated by millimolar concentrations of cysteine. Steady-state initial velocity measurements indicate that the enzyme catalyzes the transfer of sulfane sulfur by way of a double displacement mechanism with formation of a covalent enzyme-sulfur intermediate. The turnover number for the enzyme-catalyzed reaction, with thiosulfate as donor substrate and cyanide ion as the sulfur acceptor, is 260 s-1. This value corresponds to a catalytic efficiency 60% of that measured for a previously characterized bovine liver enzyme of more than twice the molecular weight. Furthermore, KmCN is 24 mM which is 2 orders of magnitude higher than the value observed previously for the bovine enzyme. Evidence from chemical inactivation studies implicates an essential sulfhydryl group in the enzyme activity. It is proposed that this group is the site of substrate-sulfur binding in the obligatory enzyme-sulfur intermediate. Furthermore, a cationic site important for binding of the donor thiosulfate is tentatively identified from anion inhibition studies. Tests of alternate acceptor substrates indicate that the physiological dithiol, dihydrolipoate, is a more efficient acceptor than cyanide ion for the enzyme-bound sulfur. Of possibly greater physiological significance, it has been found that the enzyme catalyzes the formation of iron-sulfur centers. Other work indicates the E. coli rhodanese is subject to catabolite repression and suggests a physiological role for the enzyme in aerobic energy metabolism.

Anions↗

Copper toxicosis in cattle fed chicken litter.

Cattle from 2 herds developed copper toxicosis after the ingestion of chicken litter. The affected animals were adult Holstein cows and crossbred steers that ate 9 to 16 kg of litter/day. These cattle developed a sudden onset of weakness, depression, anorexia, icteric mucous membranes, and dark reddish brown urine. Liver copper concentrations in 2 cattle (1 from each herd) were 436 and 730 ppm. Results of copper analyses of chicken litter ranged from 620 to 920 ppm. Sodium molybdate and sodium thiosulfate were added to the ration of the dairy herd. Two cows with clinical signs of copper toxicosis recovered after being given additional sodium molybdate and thiosulfate supplements, orally.

Animal Feed↗

Experimental and clinical results with intraperitoneal cisplatin.

Up to 30% of ovarian cancer patients with minimal residual disease may achieve a complete remission with intraperitoneal cisplatin therapy. This mode of therapy, however, is toxic and cumbersome. Sodium thiosulfate was shown to protect renal function, as well as lessen hematologic complications of cisplatin therapy. A major problem with cisplatin in terms of achieving maximal therapeutic dosages is neurotoxicity. Different platinum chemotherapeutic agents, pharmacodynamics, and pharmacokinetics of intraperitoneal administration, and rescue agents other than sodium thiosulfate are presently being investigated by The Netherlands Cancer Institute.

Antineoplastic Combined Chemotherapy Protocols↗

[Glycerin metabolism in purple sulfur bacteria].

The purple sulfur bacteria Thiocapsa roseopersicina, strains BBS and SL-4, and Lamprobacter modestohalophilus assimilate glycerol when grown in the light under the anaerobic conditions or in the dark under the aerobic conditions. Apart from glycerol, the bacteria require for their growth reduced sulfur compounds (thiosulfate) and bicarbonate. The 14C from 1-14C-glycerol is incorporated into the cells of T. roseopersicina and L. modestohalophilus at a lower rate in the presence of thiosulfate under the phototrophic conditions. In the presence of glycerol, all the studied purple sulfur bacteria produce more reserve carbohydrates of the glucan type than on the mineral medium. Glycerol metabolism in purple sulfur bacteria, like that in nonsulfur purple bacteria, involves glycerokinase and the pyridine nucleotide independent alpha-glycerophosphate dehydrogenase. The activity of these enzymes does not depend on the conditions of the cultural growth and the presence of glycerol in the medium.

Anaerobiosis↗

[Peroxidative and antioxidative activity of lipids in experimental pancreatitis].

Peroxidation of lipids and alteration in antioxidative activity were studied in normal state and in experimental pancreatitis within 1, 3, 7, 14 and 30 days after the operation. Experimental pancreatitis was accompanied by increase in content of lipid peroxides and by decrease in antioxidative activity of lipids extracted from liver tissue. Administration of thiosulfate decreased slightly the lipid peroxides content and increased the level of antioxidants in pancreatitis. The data obtained suggest that thiosulfate is effective inhibitor of free radical oxidation; it may be used as therapeutic drug in acute pancreatitis.

Animals↗

Effects of various treatments on induced chronic aflatoxicosis in rabbits.

Male New Zealand White rabbits were orally given 0.05 mg of aflatoxin B1 (AFB1)/kg of body weight daily for 10 days and were treated with glutathione-precursors and depletor, antibacterial agents, or sodium thiosulfate. The drug administered, the mortality, and the mean survival time were as follows: corn-oil controls (0), euthanatized at 25 days; AFB1-controls (2), 21 days; AFB1 and saline controls (2), 22 days; cysteine and AFB1 (5), 13 days; methionine and AFB1 (5), 12 days; sodium thiosulfate and AFB1 (2), 21 days; sulfadimethoxine and AFB1 (1), 24 days; oxytetracycline and AFB1 (0), euthanatized at 25 days; and ethyl maleate and AFB1 (3), 21 days. Clinical signs of toxicosis included decreased feed consumption during AFB1 administration, loss of body weight or failure to gain, and death. Clinicopathologic changes included increases in serum bilirubin concentration and alanine aminotransferase and aspartate aminotransferase activities. Prothrombin and activated partial thromboplastin times were lengthened. Plasma fibrinogen concentration was decreased. Changes in PCV, hemoglobin concentration, and serum alkaline phosphatase were unremarkable. Oxytetracycline had protective effects against chronic aflatoxicosis in rabbits. Cysteine and methionine enhanced chronic aflatoxicosis.

Aflatoxin B1↗

Stability and inactivation of mutagenic drugs and their metabolites in the urine of patients administered antineoplastic therapy.

Urine samples from patients administered mutagenic antineoplastic drugs are mutagenic in the Ames assay, and hence may pose a genotoxic hazard to hospital personnel or family members caring for the patient. The urine samples in the present study were tested for mutagenicity in several strains of Salmonella typhimurium that were uvr negative (TA98, TA100) or positive (TA102, UTH8413, UTH8414), and were analyzed for the presence of drugs and their metabolites using high-pressure liquid chromatography (HPLC). Urine samples from cancer patients were kept at room temperature and their mutagenicity as well as the chemical stability of the drugs was tested for a period of 14 days. It was observed that, in general, the urine remained mutagenic for the 14-day period while the parent compound degraded within the first seven days. An exception was cisplatin, which was chemically stable as platinum, but the urine decreased in mutagenicity with time. This decrease was probably the result of ligand exchange with the platinum. Inactivation methods were developed to reduce the genotoxic hazard posed by the mutagenic compounds in the urine. Cisplatin was inactivated by complexing with sodium diethyldithiocarbamate (DDTC). Oxidation of urine containing mitomycin C and doxorubicin (sodium thiosulfate must be added to urine containing doxorubicin) with 5.25% sodium hypochlorite solution (bleach) results in mutagenic inactivation. Urine containing cyclophosphamide and its metabolites was oxidized with alkaline potassium permaganate and the active degradation products trapped with sodium thiosulfate. Both chemical and mutagenic assays are necessary to determine the reduction of risk. Methods of inactivation of mutagenic urine developed in this study are both effective and practical for the reduction of exposure to genotoxic hazards.

Antineoplastic Agents↗

Action of gold sodium thiomalate on experimental thrombosis in vivo.

In order to study the effect of gold compounds on the action of thrombin in vivo, experiments were performed to measure platelet survival and the weight of thrombus formation in experimental models of intra-aortic thrombosis by two indwelling aortic catheter methods. We have called these the long and short catheter methods. Platelet survival was reduced in all gold-treated and control animals which had indwelling aortic catheters. In the long catheter model, New Zealand White male rabbits were treated with one of the following: gold sodium thiomalate, sterile water, gold thioglucose, gold sodium thiosulfate, disodium thiomalate. Gold sodium thiomalate-treated rabbits had a reduced weight of experimentally induced intra-aortic thrombi compared with animals treated with sterile water or equimolar concentrations of gold thioglucose, gold sodium thiosulfate, or disodium thiomalate. This reduction in thrombus weight in the animals treated with gold sodium thiomalate was not reflected by changes in platelet survival or fibrinolysis. The serum gold levels achieved in these in vivo experiments was in the range of 5.0 X 10(-5) to 1.0 X 10(-4) M. These values are comparable to levels which can be achieved in human subjects immediately after a gold injection. In the short catheter model, New Zealand White male rabbits were treated with either gold sodium thiomalate, gold thioglucose, disodium thiomalate, or auranofin. Controls were given either water or 0.05% chlorocresol. Water-treated and gold sodium thiomalate-treated animals were also given 51Cr-labeled platelets and 125I-fibrinogen before insertion of the catheter.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Capillary electrophoretic separation of inorganic sulfur-sulfide, polysulfides, and sulfur-oxygen species.

A CE protocol was developed for the identification and separation of inorganic polysulfides simultaneously with other inorganic sulfur-bearing species coexisting in aqueous hydrosulfide/sulfur solutions. The electrophoretic separation of thiosulfate, sulfate, hydrosulfide, sulfite, tetrathionate, and polysulfides was achieved at pH values between 8.2 and 12.2. The peaks attributed to the polysulfide species were strongly sensitive to pH. CE analysis of hydrosulfide/sulfur solutions at different pH values permitted possible identification of two forms of polysulfides: S4(2-) and S3(2-). Upon exposure to air at ambient temperature, thiosulfate was the main oxidation product of hydrosulfide/sulfur solutions mainly in the first 60 min, when hydrosulfide was rapidly consumed. Analysis of the oxidation reaction products provided retrospectively tentative evidence that the peaks separated and identified as tri- and tetrasulfide may be ascribed to polysulfides.

Electrophoresis, Capillary↗

Developmental toxicology of acetonitrile in the Syrian golden hamster.

Pregnant hamsters were exposed to acetonitrile by inhalation, ingestion, or ip injection during the early primitive streak stage of embryogenesis. Inhalation of 1,800 or 3,800 ppm acetonitrile for 60 min failed to induce malformations in the near-term offspring whereas inhalation of 5,000 or 8,000 ppm acetonitrile was associated with production of severe axial skeletal (dysraphic) disorders. One fetus afflicted with extrathoracic ectopia cordis was recovered from a dam exposed to 8,000 ppm acetonitrile. An oral or ip dose of 100-400 mg/kg acetonitrile in hamsters of equivalent gestational age also caused malformations identical to those noted following inhalation exposure. Some dams exposed to the highest concentrations or doses of acetonitrile displayed overt signs of poisoning. Multiple injections of sodium thiosulfate antagonized the mortality and signs of intoxication associated with acetonitrile treatment. Offspring of thiosulfate-treated hamsters exposed to acetonitrile failed to exhibit the marked teratogenic response that was associated with exposure to equivalent concentrations or doses of acetonitrile alone. Elevated concentrations of cyanide and thiocyanate were detected in all tissues studied at 2.5 hr after an oral or ip dose of acetonitrile. Cyanide was liberated when acetonitrile was incubated in vitro with hamster liver slices or NADPH-fortified hepatic microsomal preparations. The results suggested that in vivo liberation of cyanide from acetonitrile was responsible for the production of terata.

Abnormalities, Drug-Induced↗

High-dose biweekly intraperitoneal cisplatin: an effective way to increase cisplatin dose intensity.

The amount of cisplatin (DDP) delivered per unit time (dose intensity, expressed in mg/m2/week) may be an important factor in determining the clinical outcome in tumors such as ovarian carcinoma. In this neoplasm, intraperitoneal chemotherapy is an effective form of treatment. In this trial we have explored the tactic of shortening the cycle interval as a way to increase the dose intensity of ip DDP. Sixteen patients with a variety of solid tumors received a total of 77 cycles of DDP 180 mg/m2 instilled ip concurrent with i.v. sodium thiosulfate at the dose of 4 g/m2 as loading dose, followed by 12 g/m2 over 6 hr. Each cycle was repeated every 2 weeks. The number of cycles delayed for 3 or more days was 28 (36%). The mean DDP dose intensity received by these patients was 77% of the planned dose or 69 mg/m2/week (confidence interval 95%, 60.5-77.5). The treatment was generally well tolerated: myelotoxicity was mild, only 1 patient had an increase in serum creatinine to > 2 mg/dl. Five patients (31%) developed symptoms of peripheral neuropathy. All patients were evaluable for response. The overall response rate (complete plus partial) in these heavily pretreated patients was 19%. When DDP is given in high doses by the ip route concurrently with systemic sodium thiosulfate, the dosing interval can be reduced to every 2 weeks permitting a marked increase in DDP dose intensity.

Abdominal Neoplasms↗

Anaerobic degradation of 1,3-propanediol by sulfate-reducing and by fermenting bacteria.

Three strains of strictly anaerobic Gram-negative, non-sporeforming, motile bacteria were enriched and isolated from freshwater sediments with 1,3-propanediol as sole energy and carbon source. Strain OttPdl was a sulfate-reducing bacterium which grew also with lactate, ethanol, propanol, butanol, 1,4-butanediol, formate or hydrogen plus CO2, the latter only in the presence of acetate. In the absence of sulfate, most of these substrates were fermented to the respective fatty acids in syntrophic cooperation with Methanospirillum hungatei. Sulfur, thiosulfate, or sulfite were reduced, nitrate not. The other two isolates degraded propanediol only in coculture with Methanospirillum hungatei. Strain OttGlycl grew in pure culture with acetoin and with glycerol in the presence of acetate. Strain WoAcl grew in pure culture only with acetoin. Both strains did not grow with other substrates, and did not reduce nitrate, sulfate, sulfur, thiosulfate or sulfite. The isolates were affiliated with the genera Desulfovibrio and Pelobacter. The pathways of propanediol degradation and the ecological importance of this process are discussed.

Anaerobiosis↗

Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. I. Isolation of new sulfate-reducing bacteria enriched with acetate from saline environments. Description of Desulfobacter postgatei gen. nov., sp. nov.

Three strains (2ac9, 3ac10 and 4ac11) of oval to rod-shaped, Gram negative, nonsporing sulfate-reducing bacteria were isolated from brackish water and marine mud samples with acetate as sole electron donor. All three strains grew in simple defined media supplemented with biotin and 4-aminobenzoic acid as growth factors. Acetate was the only electron donor utilized by strain 2ac9, while the other two strains used in addition ethanol and/or lactate. Sulfate served as electron acceptor and was reduced to H2S. Complete oxidation of acetate to CO2 was shown by stoichiometric measurements with strain 2ac9 in batch cultures using sulfate, sulfite or thiosulfate as electron acceptors. With sulfate an average growth yield of 4.8 g cell dry weight was obtained per mol of acetate oxidized; with sulfite or thiosulfate the growth yield on acetate was about twice as high. None of the strains contained desulfoviridin. In strain 2ac9 cytochromes of the b- and c-type were detected. Strain 2ac9 is described as type strain of the new species and genus, Desulfobacter postgatei.

Acetates↗

O-Acetylserine sulfhydrylase and S-sulfocysteine synthase activities of Rhodospirillum tenue.

O-Acetylserine sulfhydrylase in cell-free extracts of Rhodospirillum tenue was markedly repressed after growth in the presence of sulfide or thiosulfate, whereas S-sulfocysteine synthase activity remained almost unchanged. Purification on DE52 cellulose resulted in the separation of two proteins: Protein I with a molecular weight of 57000 had O-acetylserine sulfhydrylase activity only, while protein II with a molecular weight of 46000 had S-sulfocysteine synthase activity in addition. The activity of protein II with O-acetylserine plus sulfide was about 1.5 of that with O-acetylserine plus thiosulfate. Protein I from sulfate-grown cells possessed 74% of the total O-acetylserine sulfhydrylase, protein II 26%. Growth with sulfide repressed only the synthesis of protein I, which after separation showed only 19% of the measurable O-acetylserine sulfhydrylase, whereas protein II now possessed 81%. Regulatory and kinetic phenomena of the two activities were studied. In addition to the phototrophic bacteria studied earlier, also Rhodomicrobium vannielii, Rhodopseudomonas acidophila, Rhodocyclus purpureus and Thiocystis violacea were found to contain O-acetylserine sulfhydrylase activities; the latter two species contained S-sulfocysteine synthase activities in addition.

Cysteine↗

A new staining method of astrocytes for paraffin section.

A new method of staining astrocytes in formation-fixed, paraffin-embedded sections was devised: (1) fix them in 5% mercuric chloride solution for 30 min to 1 h at 56 degrees C, (2) then place in 0.5% iodine alcohol for 5 min followed by placing in 0.5% sodium thiosulfate for 5 min, (3) immerse in 0.25% potassium permanganate for 3 min, (4) place in 2% oxalic acid for 2 min. (5) mordant in 2% iron alum for 45 s, and (6) place in 2% silver nitrate solution for 30 min. The next step is impregnation in ammoniacal silver solution for 10 -- 15 min at 56 degrees C, followed by reduction in neutral formalin and 2% iron alum, toning in 0.2% gold chloride, and fixing in 5% sodium thiosulfate. Pathological astrocytes of fibrillary and protoplasmic types were distincly demonstrated, although nerve cells and nuclei of oligodendrocytes and microglial cells were also faintly stained. Thus, the staining for paraffin sections is fairly selective for astrocytes in pathological states.

Astrocytes↗